Exposure of broiler chickens to deoxynivalenol and Campylobacter jejuni induces substantial changes in intestinal gene expression

The mycotoxin deoxynivalenol (DON) is of high importance among feed contaminants because of its frequent occurrence in toxicologically relevant concentrations worldwide. Cereal crops, the main component of chicken diet, are commonly contaminated with DON, resulting in frequent exposure of chickens to DON. Likewise, Campylobacter (C.), a pathogen of major public and animal health concern, is frequently found in chicken flocks and poses a threat to the One Health approach. Campylobacter colonizes the gastrointestinal (GI) tract of poultry with a high bacterial load in the caeca. However, the mechanism of C. jejuni colonization in chickens is still not understood albeit it is well known that C. jejuni resides primarily in the mucosal layer of the chicken intestine. Therefore, in the actual study we focused on the effect of exposure to DON and/or C. jejuni on expression profiles of intestinal mucins (MUC1, MUC2), beta-defensins (Gallinacin (GAL) 10, 12), cytokines (Toll-like receptor 2 (TLR2), Interleukin (IL) 6, 8, Interferon-gamma (IFN)-gamma), inducible nitric oxide synthase 2 (iNOS2), as well as selected tight junction proteins (Claudin 5 (CLDN5), Occludin (OCLN), and zonula occludens-1 (ZO1) via RT-qPCR. For this, a total of 150 one-day-old Ross 308 broiler chickens were randomly allocated to six different groups (n = 25 with 5 replicates/group) and were fed for 5 weeks with either contaminated diets (5 or 10 mg DON/kg feed) or basal diets (control). Following oral infection of birds with C. jejuni NCTC 12744 at 14 days of age, several changes in gene expression patterns were demonstrated. A significant (P <= 0.05) downregulation of MUC2 mRNA expression was observed in birds fed DON5 and DON10 diet, as well as in birds co-exposed to DON5 and C. jejuni at 7 dpi. Furthermore, at 14 dpi, MUC2 mRNA expression was significantly (P <= 0.05) downregulated in birds fed DON (5 mg and 10 mg/kg diet) with and without C. jejuni and in birds infected solely with C. jejuni. The actual study also demonstrated that co-exposure of broiler chickens to DON and C. jejuni resulted in a decreased barrier function via downregulation of OCLD mRNA expression. In addition, Campylobacter infection induced an increased expression of the antimicrobial peptide GAL12 and the IL8 gene, indicating that C. jejuni can initiate an immune response in the chicken gut in a proinflammatory manner. Similarly, DON with and without C. jejuni induced upregulation of GAL10 and GAL12 mRNA expression at 7 dpi. Moreover, no change in iNOS2 mRNA expression was observed in both the jejunum and the cecum at either 7 dpi or 14 dpi, suggesting unchanged NO production during exposure/infection. In conclusion, we confirmed that DON contamination corresponding to the currently applicable EU guidance value of 5 mg DON/kg feed affects the intestinal gene expression profiles of broilers, mainly in a dose-independent manner. Furthermore, DON exposure interacted synergistically with C. jejuni challenge regarding mucins, innate immunity gene expression in either the jejunum or the cecum, suggesting immunomodulatory activity of both foodborne agents (DON and C. jejuni). The mycotoxin deoxynivalenol (DON) is of high importance among feed contaminants because of its frequent occurrence in toxicologically relevant concentrations worldwide. Cereal crops, the main component of chicken diet, are commonly contaminated with DON, resulting in frequent exposure of chickens to DON. Likewise, Campylobacter (C.), a pathogen of major public and animal health concern, is frequently found in chicken flocks and poses a threat to the One Health approach. Campylobacter colonizes the gastrointestinal (GI) tract of poultry with a high bacterial load in the caeca. However, the mechanism of C. jejuni colonization in chickens is still not understood albeit it is well known that C. jejuni resides primarily in the mucosal layer of the chicken intestine. Therefore, in the actual study we focused on the effect of exposure to DON and/or C. jejuni on expression profiles of intestinal mucins (MUC1, MUC2), β-defensins (Gallinacin (GAL) 10, 12), cytokines (Toll-like receptor 2 (TLR2), Interleukin (IL) 6, 8, Interferon-γ (IFN)-γ), inducible nitric oxide synthase 2 (iNOS2), as well as selected tight junction proteins (Claudin 5 (CLDN5), Occludin (OCLN), and zonula occludens-1 (ZO1) via RT-qPCR. For this, a total of 150 one-day-old Ross 308 broiler chickens were randomly allocated to six different groups (n = 25 with 5 replicates/group) and were fed for 5 weeks with either contaminated diets (5 or 10 mg DON/kg feed) or basal diets (control). Following oral infection of birds with C. jejuni NCTC 12744 at 14 days of age, several changes in gene expression patterns were demonstrated. A significant (P ≤ 0.05) downregulation of MUC2 mRNA expression was observed in birds fed DON5 and DON10 diet, as well as in birds co-exposed to DON5 and C. jejuni at 7 dpi. Furthermore, at 14 dpi, MUC2 mRNA expression was significantly (P ≤ 0.05) downregulated in birds fed DON (5 mg and 10 mg/kg diet) with and without C. jejuni and in birds infected solely with C. jejuni. The actual study also demonstrated that co-exposure of broiler chickens to DON and C. jejuni resulted in a decreased barrier function via downregulation of OCLD mRNA expression. In addition, Campylobacter infection induced an increased expression of the antimicrobial peptide GAL12 and the IL8 gene, indicating that C. jejuni can initiate an immune response in the chicken gut in a proinflammatory manner. Similarly, DON with and without C. jejuni induced upregulation of GAL10 and GAL12 mRNA expression at 7 dpi. Moreover, no change in iNOS2 mRNA expression was observed in both the jejunum and the cecum at either 7 dpi or 14 dpi, suggesting unchanged NO production during exposure/infection. In conclusion, we confirmed that DON contamination corresponding to the currently applicable EU guidance value of 5 mg DON/kg feed affects the intestinal gene expression profiles of broilers, mainly in a dose-independent manner. Furthermore, DON exposure interacted synergistically with C. jejuni challenge regarding mucins, innate immunity gene expression in either the jejunum or the cecum, suggesting immunomodulatory activity of both foodborne agents (DON and C. jejuni).© 2025. The Author(s).

DSM Austria GmbH
Biomin Holding GmbH, Research Center
LVA GmbH
Veterinärmedizinische Universität Wien